NGS Library Preparation

Illumina Libraries

RNA-Seq Kits

10X Genomics


PacBio SMRTbell Libraries

SMRTbell library preparation does not utilize amplification steps; the completed library molecules are used as direct templates for the sequencing process.  Due to this, the quality of the starting DNA has enormous influence on the success and quality of the sequencing results.  High-quality, high-molecular-weight genomic DNA is extremely important for obtaining long reads and optimal performance.

Sample Submission Requirements:

  • Gel image submitted with sample showing no signs of degradation.  For amplicon or cDNA samples, a Bioanalyzer trace can be submitted instead.
  • Nanodrop readings: 
    • 260/280: ratio should be around ~1.8; higher values are acceptable, low values indicate contamination/very low concentration.
    • 260/230: ratio should be ~2.0-2.2
  • Sample Concentration:
    • Use Qubit or PicoGreen for best results
    • Submit a minimum of 1.5 ug for <2kb libraries, 2.5 ug for 5 kb libraries, 15 ug for 20kb+
  • Volume and Buffers:
    • Max volume 130 uL
    • Avoid buffers using EDTA for DNA resuspension
  • Additonal Best Practices can be found here